LTB4 synthesis inhibitors

ABSTRACT

This invention relates to a compound of the formula: ##STR1## or a pharmaceutically acceptable salt thereof wherein X is oxygen, sulfur, --CH═CH--, or --CH═N--; 
     wherein R 1  is --CO 2  R 2  or tetrazole; 
     wherein R 2  is hydrogen, alkyl of 1 to 6 carbons or a pharmaceutically acceptable cation; 
     wherein R is an alkyl of from 1 to 20 carbons, --(CH 2 ) p  CF 3  or --(CH 2 ) q  R 3  wherein R 3  is alkoxy, phenoxy or alkoxy substituted phenoxy wherein the alkoxy group has from 1 to 8 carbons; 
     wherein p and q are integers from 0 to 20; 
     wherein n is 0 or 1; and 
     wherein m is 0, 1, 2, or 3.

This is a CONTINUATION of prior application Ser. No. 07/707,522 filed onMay 30, 1991 now U.S. Pat. No. 5,356,919.

BACKGROUND OF THE INVENTION

The present invention relates to pharmaceutical agents (compounds) whichact as leukotriene B₄ (LTB₄) synthesis inhibitors in mammals. Thecompounds inhibit LTB₄ synthesis by inhibiting phospholipase A₂ (PLA₂)activity. PLA₂ is an important enzyme in the biosynthesis ofleukotrienes as PLA₂ acts to release arachidonic acid fromphospholipids. Once released, arachidonic acid is rapidly metabolized bya variety of enzymes of the arachidonic acid cascade to produceprostaglandins, leukotrienes and related compounds. The use of thecompounds herein to inhibit PLA₂ activity thus inhibits the release ofarachidonic acid from phospholipids. The inhibition of release ofarachidonic acid similarly diminishes subsequent products in thearachidonic acid cascade, such as prostaglandins, leukotrienes, andrelated compounds, including LTB₄.

LTB₄ (Formula I) is an arachidonic acid metabolite which is produced bythe 5-lipoxygenase pathway. Pharmacologically, LTB₄ is an importantmediator of ##STR2## inflammation. LTB₄ is known to induce chemotaxis,chemokinesis, aggregation, and degranulation of leukocytes in vitro, andto induce accumulation of polymorphonuclear leukocytes, and increasevascular permeability and edema formation in vivo. Particularly highlevels of LTB₄ are detected in lesions in inflammatory diseases such asrheumatoid or spondylarthritis, gout, psoriasis, ulcerative colitis,Crohn's disease, multiple sclerosis and some respiratory diseases. Sincethe compounds herein inhibit PLA₂ and thereby LTB₄ synthesis, thecompounds of the present invention are useful in treating inflammatoryconditions in mammals such as psoriasis, Crohn's disease, ulcerativecolitis, multiple sclerosis and the like.

Accordingly, it is an object of this invention to produce compounds foruse as pharmaceutical agents which will exhibit LTB₄ inhibitory activityin mammals.

The pharmacology of the biologically active leukotrienes is generallydiscussed in J. Clin. Invest. 73, 889-897 (1984).

SUMMARY OF THE INVENTION

This invention relates to a compound of the formula: ##STR3## or apharmaceutically acceptable salt thereof wherein X is oxygen, sulfur,--CH═CH--, or --CH═N--;

wherein R¹ is --CO₂ R² or tetrazole;

wherein R² is hydrogen, alkyl of 1 to 6 carbons or a pharmaceuticallyacceptable cation;

wherein R is an alkyl of from 1 to 20 carbons, --(CH₂)_(p) CF₃ or--(CH₂)_(q) R³ wherein R³ is alkoxy, phenoxy or alkoxy substitutedphenoxy wherein the alkoxy group has from 1 to 8 carbons;

wherein p and q are integers from 0 to 20;

wherein n is 0 or 1; and

wherein m is 0, 1, 2, or 3.

This invention, more specifically, relates to a compound of the formula:##STR4## or a pharmaceutically acceptable salt thereof wherein X isoxygen, sulfur, --CH═CH--, or --CH═N--;

wherein R¹ is --CO₂ R² or tetrazole;

wherein R² is hydrogen, alkyl of 1 to 6 carbons or a pharmaceuticallyacceptable cation;

wherein R is an alkyl of from 1 to 20 carbons, --(CH₂)_(p) CF₃ or--(CH₂)_(q) R³ wherein R³ is alkoxy, phenoxy or alkoxy substitutedphenoxy wherein the alkoxy group has from 1 to 8 carbons;

wherein p and q are integers from 0 to 20;

wherein n is 0 or 1; and

wherein m is 0, 1, 2, or 3.

This invention also relates to a compound of the formula: ##STR5## or apharmaceutically acceptable salt thereof wherein X is oxygen, sulfur,--CH═CH--, or --CH═N--;

wherein R¹ is --CO₂ R² or tetrazole;

wherein R² is hydrogen, alkyl of 1 to 6 carbons or a pharmaceuticallyacceptable cation;

wherein R is an alkyl of from 1 to 20 carbons, --(CH₂)_(p) CF₃ or--(CH₂)_(q) R³ wherein R³ is alkoxy, phenoxy or alkoxy substitutedphenoxy wherein the alkoxy group has from 1 to 8 carbons;

wherein p and q are integers from 0 to 20;

wherein n is 0 or 1; and

wherein m is 0, 1, 2, or 3.

This invention also relates to a compound of the formula: ##STR6## or apharmaceutically acceptable salt thereof wherein X is oxygen, sulfur,--CH═CH--, or --CH═N--;

wherein R¹ is --CO₂ R² or tetrazole;

wherein R² is hydrogen, alkyl of 1 to 6 carbons or a pharmaceuticallyacceptable cation;

wherein R is an alkyl of from 1 to 20 carbons, --(CH₂)_(p) CF₃ or--(CH₂)_(q) R³ wherein R³ is alkoxy, phenoxy or alkoxy substitutedphenoxy wherein the alkoxy group has from 1 to 8 carbons;

wherein p and q are integers from 0 to 20;

wherein n is 0 or 1; and

wherein m is 0, 1, 2, or 3.

DETAILED DESCRIPTION

This invention encompasses compounds of Formulae II-V as previouslydescribed. A particularly preferred embodiment of the present inventionis encompassed by a compound of the formula: ##STR7## or apharmaceutically acceptable salt thereof wherein X is oxygen, sulfur,--CH═CH--, or --CH═N--;

wherein R¹ is --CO₂ R² or tetrazole;

R² is hydrogen, alkyl of 1 to 6 carbons or a pharmaceutically acceptablecation;

wherein R is an alkyl of from 1 to 20 carbons, --(CH₂)_(p) CF₃ or--(CH₂)_(q) R³ wherein R³ is alkoxy, phenoxy or alkoxy substitutedphenoxy wherein the alkoxy group has from 1 to 8 carbons;

wherein p and q are integers from 0 to 20;

wherein n is 0 or 1; and

wherein m is 0, 1, 2, or 3.

The term "lower alkyl" as used herein means straight or branched chainalkyls having 1-6 carbon atoms.

The term "pharmaceutically acceptable cation" as used to describe R²refers to cations such as ammonium, sodium, potassium, lithium, calcium,magnesium, ferrous, zinc, copper, manganous, aluminum, ferric, manganic,ammonium, tetraalkyl-ammonium, and the like.

The term "pharmaceutically acceptable salts" refers either to thosenon-toxic, base derived salts of any compound herein having a carboxylicacid function.

The base derived salts can be derived from pharmaceutically acceptablenon-toxic inorganic or organic bases. Among the inorganic bases employedto produce pharmaceutically acceptable salts are the hydroxide bases ofthe "pharmaceutically acceptable cations" disclosed above.

Among the organic bases employed to produce pharmaceutically acceptablesalts are the pharmaceutically acceptable non-toxic bases of primary,secondary, and tertiary amines. Especially preferred non-toxic bases areisopropylamine, diethylamine, ethanolamine, dicyclohexylamine, choline,and caffeine.

All of the pharmaceutically acceptable salts are prepared byconventional processes which are well known to those of ordinary skillin the art.

The compounds of this invention are generally prepared according to thereaction schemes I, II and III, wherein a side chain is substituted ontoa halo aromatic acid or ester moiety. By halo is meant a halogen such asbromo, iodo or chloro. In Scheme I, the halo group is represented by theterm "halo." By aromatic moiety is meant phenyl, pyridyl, thienyl orfuryl, corresponding to "X" in the aryl ring being --CH═CH--, --CH═N--,--S--, and --O--.

As disclosed in the following reaction Schemes I-IV, an alkyne sidechain can be added to an aromatic moiety by different techniques. Thealkyne side chain is then hydrated with sulfuric acid, water andmercuric oxide to yield the indicated diketo product.

In Scheme I the alkyne side chain is added by performing a nucleophilicsubstitution of the halogen such as via a coupling reaction with analkyne, CO, and Pd[O]. In Scheme II the alkyne side chain is added byacylation of a trimethylsilyl (TMS) acetylide with a diacid chloride inthe presence of AlCl₃. In Scheme III the alkyne side chain is added vianucleophilic attack of an acetylide anion on an aldehyde with subsequentoxidation of the resulting alcohol. In Scheme IV the alkyne side chainis added via nucleophilic attack of an acetylide anion on an acidchloride.

The biological activity possessed by the compounds of this invention wasindicated by positive results in assays for inhibition of human synovialfluid PLA₂ (HSF-PLA₂) and LTB₄ biosynthesis in HL-60 cells.

By virtue of their activity as LTB₄ synthesis inhibitors, the compoundsof Formula I-VI are useful in treating inflammatory conditions inmammals such as psoriasis, Crohn's disease, ulcerative colitis, multiplesclerosis and the like. Similarly, the compounds of Formula I-VI can beused in preventing recurring inflammatory attacks. A physician orveterinarian of ordinary skill can readily determine whether a subjectexhibits the inflammatory condition. The preferred utility relates totreatment of ulcerative colitis.

The compounds of the present invention can be administered in such oraldosage forms as tablets, capsules, soft gels, pills, powders, granules,elixirs, or syrups.

The compounds can also be administered intravascularly,intraperitoneally, subcutaneously, intramuscularly, or topically usingforms known to the pharmaceutical art. Moreover, they can beadministered rectally or vaginally, in such forms as suppositories orbougies. In general, the preferred form of administration is oral. Forthe orally administered pharmaceutical compositions and methods of thepresent invention, the foregoing active ingredients will typically beadministered in admixture with suitable pharmaceutical diluents,excipients, or carriers (collectively referred to herein as "carrier"materials) suitably selected with respect to the intended form ofadministration, that is, oral tablets, capsules, soft gels, elixirs,syrups, drops, and the like, and consistent with conventionalpharmaceutical practices.

For example, for oral administration in the form of tablets or capsules,a therapeutically effective amount of one or more compounds of thepresent invention can be combined with any oral non-toxicpharmaceutically acceptable inert carrier such as lactose, starch,sucrose, cellulose, magnesium stearate, dicalcium phosphate, calciumsulfate, mannitol, and the like, or various combinations thereof. Fororal administration in liquid forms, such as in soft gels, elixirs,syrups, drops and the like, a therapeutically effective amount of theactive drug components can be combined with any oral non-toxicpharmaceutically acceptable inert carrier such as water, saline,ethanol, polyethylene glycol, propylene glycol, corn oil, cottonseedoil, peanut oil, sesame oil, benzyl alcohol, various buffers, and thelike, or various combinations thereof. Moreover, when desired ornecessary, suitable binders, lubricants, disintegrating agents, andcoloring agents can also be incorporated in the mixture. Suitablebinders include starch, gelatin, natural sugars, corn sweeteners,natural and synthetic gums such as acacia, sodium alginate,carboxymethylcellulose, polyethylene glycol, and waxes, or combinationsthereof. Lubricants for use in these dosage forms include boric acid,sodium benzoate, sodium acetate, sodium chloride, and the like, orcombinations thereof. Disintegrators include, without limitation,starch, methylcellulose, agar, bentonite, guar gum, and the like, orcombinations thereof. Sweetening and flavoring agents and preservativescan also be included where appropriate.

For intravascular, intraperitoneal, subcutaneous, or intramuscularadministration, one or more compounds of the present invention can becombined with a suitable carrier such as water, saline, aqueousdextrose, and the like. For topical administration, such as forpsoriasis, therapeutically effective amounts of one or more compounds ofthe present invention can be combined with pharmaceutically acceptablecreams, oils, waxes, gels and the like. Regardless of the route ofadministration selected, the compounds of the present invention areformulated into pharmaceutically acceptable dosage forms by conventionalmethods known to those skilled in the art. The compounds can also beformulated using pharmacologically acceptable base addition salts.Moreover,the compounds or their salts may be used in a suitable hydratedform.

Regardless of the route of administration selected, a non-toxic buttherapeutically effective quantity of one or more compounds of thisinvention is employed in any treatment. The dosage regimen forpreventing or treating inflammatory conditions with the compounds ofthis invention is selected in accordance with a variety of factors,including the type, age, weight, sex, and medical condition of thepatient, the severity of the inflammatory condition, the route ofadministration, and the particular compound employed in the treatment. Aphysician or veterinarian of ordinary skill can readily determine andprescribe the effective amount of the drug required to prevent or arrestthe progress of the condition. In so proceeding, the physician orveterinarian could employ relatively low doses at first and sequentiallyincrease the dose until a maximum response is obtained. Daily dosages ofthe compounds of the invention are ordinarily in the range of about 1.0mg/kg up to about 30.0 mg/kg, (preferably in the range of about 2.0 to14.0 mg/kg (orally)).

The following examples illustrate the methods used to prepare thecompounds of this invention. These examples are given by way ofillustration only and are not meant to be construed as limiting theinvention in spirit or in scope, as many modifications in materials andmethods will be apparent from this disclosure to those skilled in theart.

In the structures herein a bond drawn across a bond in a ring indicatesthat the bond can be to any available carbon atom of the ring structure.##STR8##

EXAMPLE 1 ##STR9##

The above acid chloride was prepared from terphthalic acid by reacting0.5 g (3 mmoles) of terphthalic acid with 2 cc of [COCl]₂ (23.6 mmoles)in 10 cc of benzene and with one drop of dimethylformamide. The reagentswere mixed and warmed to 60° C. for twenty-four hours. The reactionmixture was cooled to room temperature and the volatile components wereremoved in vacuo to give the above compound as a pale yellow solid.

EXAMPLE 2

    CH.sub.3 (CH.sub.2).sub.11 C.tbd.C--TMS

The above compound was prepared by reacting an acetylene of the formulaCH₃ (CH₂)₁₁ C.tbd.CH (2.5 g, 12.87 mmoles) which was added to 25 cc oftetrahydrofuran (THF) and 50 mg of triphenylmethane (Ph₃ CH) which wasadded as an indicator. The solution was cooled to -30° C. and 1.6 molarn-butyllithium (n-BuLi) was added dropwise until the solution turnedred. Approximately 8.5 cc of n-BuLi was added. The solution was backtitrated with the acetylene compound until it became colorless. Thesolution was cooled to -78° C. and 2 cc (15.75 mmoles) of trimethylsilylchloride (TMS--Cl) was added. The solution was slowly warmed over aperiod of five hours to room temperature. The reaction was quenched withwater and extracted with hexane. The hexane was washed once with waterand once with brine and dried over magnesium sulfate (MgSO₄). Thetrimethylsilyl compound was isolated in an amount of 4.31 g (16.2mmoles).

EXAMPLE 3 ##STR10##

The above compound was prepared by reacting 3 mmoles of the acidchloride product from Example 1 with 0.8 g (3 mmoles) of theTMS-acetylene product from Example 2. The acid chloride and theTMS-acetylene product were dissolved in 10 cc of dichloromethane andcooled to 0° C. To the reaction mixture was added 0.8 g (6 mmoles) ofaluminum chloride (AlCl₃) in small portions over ten minutes. Thereaction mixture was stirred for about 1.5 hours at 0° C. The reactionwas quenched with ice and the mixture was extracted three times withdiethyl ether. The extracts were combined and washed once with water andonce with brine (saturated NaCl solution) and dried over magnesiumsulfate. Removal of the solvent in vacuo yielded 0.29 g of the aboveproduct. This was chromatographed on silica eluting with 15% ethylacetate--85% hexane. HRMS (M⁺): calculated 342.2195; found 342.2196

EXAMPLE 4 4-(1,3-dioxohexadecanyl)benzoic acid ##STR11##

The product of Example 3, 27 mg was treated with 2 ml of cold H₂ SO₄which had been cooled in an ice bath. The mixture was stirred until allof the product had dissolved. To the reaction mixture was added 1 mgHgO. The reaction mixture was maintained cool in an ice bath. Two dropsof water were added to the reaction mixture. The ice bath was removedand the reaction mixture was stirred and allowed to warm to roomtemperature for one half hour. The mixture was cooled to 0° C. andquenched with ice. The reaction mixture was diluted with water andextracted twice with ethyl acetate. The collected extracts were washedtwice with water. The washed ethyl acetate fractions were collected andthe volatile components removed and the residue chromatographed onsilica gel (gradient elution) with 5% isopropyl alcohol, 95% Hexane +1%HOAc then 10% isopropyl alcohol, 90% Hexane +1% HOAc as eluant. 0.017 gof the above product was isolated.

Analysis: Calculated for 5H₂ O: C, 71.51; H, 9.00 Found: C, 71.80; H,8.72

EXAMPLE 5 3-(1,3-dioxohexadecanyl)benzoic acid ##STR12##

The above compound was prepared by mixing 9.42 g of m-iodobenzoic acid(38 mmoles) and 8 g (38.4 mmoles) of an acetylene derivative of theformula

    H--C.tbd.C--(CH.sub.2).sub.12 --CH.sub.3

with 0.27 g (0.38 mmoles) of a palladium catalyst, pd(PPh₃)₂ Cl₂ in 100cc of diisopropyl amine. The reaction vessel was purged with carbonmonoxide. The reaction mixture was heated under a carbon monoxideatmosphere (atmospheric pressure, balloon) in an oil bath at 80° C. fortwo hours. The reaction mixture was cooled to room temperature. Thevolatile components were removed in vacuo and the residue was taken upin 5% hydrochloric acid and extracted with diethyl ether. The diethylether was washed once with 10% hydrochloric acid, twice with water, andonce with a brine solution and dried over magnesium sulfate. The solventwas removed yielding 18.65 gm of the product. A solid component wasisolated from this mixture by chromatography on silica gel (10%isopropyl alcohol, 90% hexane, 1% HOAc). Recrystallization from CH₂ Cl₂/Hexane afforded 0.66 g of the above product.

Analysis calculated: C, 73.76; H, 9.15 Found: C, 73.40; H, 9.13

EXAMPLE 6 ##STR13##

The above compound was prepared by forming a solution of 2.5 g (13.1mmoles) of a bromo-furanoic acid in 25 ml of tetrahydrofuran (THF) whichwas cooled to -78° C. To the solution was added a 1.6 molar solution ofn-butyl lithium in hexane (17.5 ml, 28 mmoles) which was stirred at -78°for one hour. Dimethylformamide (DMF) was added in an amount of 2.4 ml(30 mmoles). The solution was allowed to warm to room temperature. Thereaction mixture was quenched with water and acidified with 10%hydrochloric acid. The resultant reaction mixture was extracted twicewith ethyl acetate, the combined extracts were washed twice with waterand once with brine and subsequently dried over magnesium sulfate. Anorange solid was obtained after removal of the solvent in vacuo.Following chromatography on silica gel (gradient elution with EA-Hexanecontaining 1% HOAc) 0.73 gms. of the compound of the above formula wasobtained.

EXAMPLE 7 ##STR14##

The above compound was prepared by forming a solution of 1.25 g (6mmoles) of an acetylene of the formula H.tbd.(CH₂)₁₂ CH₃ in 25 ml. THFwhich contained 20 mg. of triphenylmethane which was added as anindicator. The solution was cooled to -50° C. and then treated with 3.75ml. (6 mmoles) of n-BuLi until the red color of the triphenylmethaneanion persisted. A few drops of the acetylene compound was added untilthe color disappeared. An amount of 0.42 gms. of the product formed inExample 6 in 10 ml. THF was added drop-wise to the solution. The mixturewas warmed to 0° C. over one-half hour. The mixture was quenched withwater and acidified with 10% hydrochloric acid. The aqueous phase wasextracted twice with ethyl acetate. The extracts were combined andwashed twice with water and once with brine and were dried overmagnesium sulfate. Chromatography on silica gel (gradient elution withEA-Hexane containing 1% HOAc) yielded 0.80 gms. of a pale yellow solid.

HRMS (M⁺) Calculated: 348.2301; Found: 348.2291.

EXAMPLE 8 ##STR15##

The compound was prepared by reacting 0.145 g. (4.2 mmoles) of theproduct from Example 7 in acetone (25 ml) and adding 1.5 g. of activatedMnO₂ portionwise over five minutes. The reaction mixture was stirred for24 hours at room temperature. The reaction mixture was poured into 10%hydrochloric acid and extracted with ethyl acetate. The extract waswashed once with water and dried over magnesium sulfate. After thesolvent was removed in vacuo a white solid remained. Followingchromatography on silica gel (gradient elution with EA-Hexane containing1% HOAc), 60 mg. of a white solid was recovered.

Analysis (for hydrate with 0.35 H₂ O) Calculated: C, 71.50; H, 8.77Found: C, 71.55; H, 8.63

EXAMPLE 9 5-(1,3-dioxohexadecanyl)-2-furancarboxylic acid ##STR16##

The product of Example 8, 22.5 mg was treated with 1 ml of cold H₂ SO₄which had been cooled in an ice bath. The mixture was stirred until allof the product had dissolved. To the reaction mixture was added 1 mgHgO. The reaction mixture was maintained cool in an ice bath. Two dropsof water was added to the reaction mixture. The ice bath was removed andthe reaction mixture was stirred and allowed to warm to room temperaturefor one half hour. The mixture was cooled to 0° and quenched with ice.The reaction mixture was diluted with water and extracted twice withethyl acetate. The collected extracts were washed twice with water. Thewashed ethyl acetate fractions were collected and the volatilecomponents removed and the residue chromatographed on silica gel(gradient elution) with 5% isopropyl alcohol, 95% Hexane +1% HOAc then10% isopropyl alcohol, 90% Hexane +1% HOAc, which yielded 0.018 g of theabove product.

Analysis: For 0.75 H₂ O Calculated: C, 66.73; H, 8.93 Found: C, 66.69;H, 8.74

EXAMPLE 10 ##STR17##

The above compound was prepared by forming a solution of 0.83 gms. (4mmoles) of an acetylene of the formula H--C.tbd.C--(CH₂)₁₂ CH₃ in 25 ml.THF containing 20 mgs. of triphenyl methane as an indicator. Thesolution was cooled to -50° C., then treated with 2.5 ml. of a 1.6 molarsolution (4 mmoles) of n-BuLi in hexane until the red color of thetriphenylmethane anion persisted. A few drops of the acetylene was addeduntil the color disappeared. The resultant lithium acetylide preparationwas cooled to -78° C. A solution (6 mmoles) of a diacid chloride of theformula prepared from the corresponding diacid and oxalyl chloride inbenzene (catalytic DMF) was cooled to -78° C. and the -78° C. solutionof lithium acetylide was added dropwise via a cannula. The reaction wasstirred for 10 minutes and quenched with water and warmed to roomtemperature. The reaction mixture was poured into water and acidifiedwith acetic acid (HOAc). The aqueous solution was extracted twice withethyl acetate and the extracts were washed twice with water, once withbrine and dried over magnesium sulfate. Following chromatography onsilica gel (gradient elution with isopropyl alcohol-hexane), 0.95 gms.of a product of the above formula was recovered.

Analysis calculated: C, 73.92; H, 8.74; N, 3.92 Found: C, 73.64; H,8.79; N, 3.86

EXAMPLE 11 6-(1,3-dioxohexadecanyl )-2-pyridinecarboxylic acid ##STR18##

The product of Example 10, 25.6 mg was treated with 2 ml of cold H₂ SO₄which had been cooled in an ice bath. The mixture was stirred until allof the product had dissolved. To the reaction mixture was added 1 mgHgO. The reaction mixture was maintained cool in an ice bath. Two dropsof water was added to the reaction mixture. The ice bath was removed andthe reaction mixture was stirred and allowed to warm to room temperaturefor one half hour. The mixture was cooled to 0° and quenched with ice.The reaction mixture was diluted with water and extracted twice withethyl acetate. The collected extracts were washed twice with water. Thewashed ethyl acetate fractions were collected and the volatilecomponents removed and the residue chromatographed on silica gel(gradient elution) with 5% isopropyl alcohol, 95% Hexane +1% HOAc then10% isopropyl alcohol, 90% Hexane +1% HOAc, which yielded 0.026 g of theabove product.

Analysis: For 0.8H₂ O Calculated: C, 67.77; H, 8.94; N, 3.59. Found: C,67.71; H, 8.53; N, 3.43.

EXAMPLE 12 ##STR19##

A compound of the above formula was prepared in the following manner. Toa cooled (0° C.) solution of 5.5 ml. (39.2 mmoles) of diisopropylaminein 50 ml. of THF was added 20.5 ml. of 1.6 molar BuLi (32.8 mmoles) tomake 32.8 mmoles of lithium diisopropylamide (LDA). The reaction mixturewas stirred for one-half hour at 0° C and cooled to -78° C. To themixture was added 2.1 g (16.4 mmoles) of 2-thiophene carboxylic acid in25 ml. THF. Additional THF was added to increase the volume to 200 ml.and the reaction mixture was stirred for one-half hour. DMF was added inan amount of 1.3 ml. (16.8 mmoles). The reaction mixture was warmed toroom temperature and stirred for 11/2 hours. The reaction mixture wasquenched with water and acidified with 1N hydrochloric acid andextracted with ethyl acetate. The organic extracts were combined anddried over magnesium sulfate. The resultant mixture was filtered andstripped to yield a yellow solid. Separation using chromatography onsilica eluting with ethyl acetate/hexane/1% acetic acid provided 1.3gms. of a yellow solid of the above formula. MP 160°-163°.

Analysis calculated: C, 46.15; H, 2.58 Found: C, 46.11; H, 2.82

EXAMPLE 13 ##STR20##

The compound with the above structure was prepared in the followingmanner. An acetylene of the formula H--C.tbd.C--(CH₂)₁₂ CH₃ in an amountof 549.3 mg. (2.6 mmoles) in 15 ml. of THF was cooled to -20°. To thesolution was added 1.6 ml. (2.6 mmoles) of n-BuLi. The reaction wasstirred for one half hour and 203.4 mg. (1.3 mmoles) of the product fromExample 13 in 10 ml. of THF was added. The mixture was stirred andmaintained at -20° for 15 minutes and allowed to warm to 0° C. andstirred for one-half hour. The reaction mixture was quenched with waterand acidified with 10% hydrochloric acid and extracted with ethylacetate. The combined organic extracts were dried over magnesiumsulfate. The extract was filtered and stripped to yield a yellow oilwhich solidified upon standing. The solid was dissolved in ethyl acetateand filtered through silica gel eluting with 100% hexane followed byethyl acetate in 1% acetic acid. The ethyl acetate fraction yielded392.8 mg. of the above compound as a yellow solid having a melting pointof 75°-90°.

Analysis calculated: C, 69.19; H, 8.85 Found: C, 69.18; H, 9.02

EXAMPLE 14 ##STR21##

A compound of the above structure was formed in the following manner.7.2 mg. (0.020 mmoles) of the compound prepared in Example 13 wasdissolved in 1 ml. acetone. To the solution was added 72 mg. (0.83mmoles) of activated manganese dioxide. The reaction mixture was stirredvigorously at room temperature overnight. The reaction mixture waspoured into 10% hydrochloric acid. The aqueous phase was extracted withethyl acetate. The organic washes were combined and dried over magnesiumsulfate. The organic phase was filtered and stripped to yield 6 mg.(0.010 mmoles) of the above product as a white solid.

HRMS (M⁺) Calculated: 362.1916 Found: 362.1910

EXAMPLE 15 5-(1,3-dioxohexadecyl)-2-thiophenecarboxylic acid ##STR22##

The compound of Example 14 (29.0 mg, 0.080 mmol) was dissolved in 2 mlof cold sulfuric acid. The reaction mixture was stirred until all of thesolid dissolved. Two drops of water were added. After stirring for 10minutes, the reaction mixture was placed in an oil bath at 60° C. for 1hour. The mixture was cooled to 0° C. and quenched with ice. The aqueousphase was extracted three times with 20 ml ethyl acetate. The combinedorganic extracts were washed with saturated NaHCO₃ and dried over MgSO₄.The resultant oil was filtered through silica gel eluting with EA/1%acetic acid to yield 14.2 mg (0.037 mmol) of the above compound as ayellow solid.

HRMS (M⁺) Calculated: 380.2022 Found: 380.1993

EXAMPLE 16 ##STR23##

A compound having the above formula was prepared in the followingmanner. Two grams (15.2 mmoles) of m-cyano-benzaldehyde along with 2.98gms. (45.8 mmoles) of NaN₃ and 2.9 gms. (21.1 mmoles) of Et₃ N.HCl, weredissolved in 50 ml. of 1-methyl-2-pyrrolidinone. The reaction mixturewas refluxed under argon. After 1 hour and 45 minutes the reactionmixture was cooled to room temperature and poured into 200 ml. of waterand acidified with 10% hydrochloric acid. The reaction mixture wasextracted with successive ethyl acetate washes. The ethyl acetateextracts were combined and washed with brine and dried over magnesiumsulfate. The ethyl acetate extract was chromatographed through silicagel, yielding 0.3 gms. of the product having the above formula as awhite solid.

EXAMPLE 17 ##STR24##

A compound of the above formula was prepared in the following manner. Asolution of 542 mg (2.6 mmoles) of an acetylene of the formulaHC.tbd.C(CH₂)₁₂ CH₃ was dissolved in 100 ml THF and cooled to -30° C. Tothe solution was added 1.6 ml. (2.6 mmoles) of a 1.6 molar n-BuLisolution which was added dropwise. The reaction mixture was stirred for15 minutes at which time 219 mg. (1.26 mmoles) of the product fromExample 16 dissolved in 8 ml. of THF was added dropwise. The solutionwas stirred and maintained at -30° C. for one-half hour, then warmed toroom temperature. The reaction mixture was quenched with water andacidified with 10% hydrochloric acid. The layers were separated and theorganic phase was washed with brine and dried over sodium sulfate. Thelayer was filtered and stripped to yield a yellow solid which uponchromatography over silica gel yielded 171.2 mg. of the above product asa white solid. M.P. 108°-110° C.

Analysis calculated: C, 72.21; H, 8.96; N, 14.65 Found: C, 72.03; H,9.00; N, 14.77

EXAMPLE 18 ##STR25##

The compound having the above formula was prepared in the followingmanner. A solution was prepared by dissolving 36.2 mg (0.095 mmoles) ofthe product from Example 17 in acetone. To the solution was added 360 mg(4.14 mmoles) of activated manganese dioxide. The reaction mixture wasstirred vigorously at room temperature overnight. The reaction mixturewas poured into 10% hydrochloric acid. The aqueous phase was extractedwith ethyl acetate and the organic washes were combined and dried overmagnesium sulfate. The organic phase was filtered and stripped to yield18 mg. of a pale yellow solid of a compound having the above formula.

Analysis calculated: C, 72.59; H, 8.48; N, 14.72 Found: C, 72.19; H,8.66; N, 14.13

HRMS (M⁺) Calculated: 380.2576 Found: 380.2579

EXAMPLE 19 1-[3-(1H-tetrazol-5-yl)phenyl]-1,3-hexadecadione ##STR26##

To 8 mg (0.021 mmol) of the compound produced in Example 18 was added 10drops of cold H₂ So₄. The reaction mixture was stirred for 5 minutes atwhich time about 1 mg of HgO was added. The reaction mixture was stirredat room temperature for 45 min. Water and ethyl acetate were added. Theorganic phase was washed with saturated NaHCO₃. The organic phase wascollected and dried over MgSO₄. The organic phase was filtered andvacuum stripped to yield 7.6 mg of white solid.

HRMS (M⁺) Calculated: 398.2682 Found: 398. 2683

EXAMPLE 20 ##STR27##

The above compound was prepared by reacting 2 g of phenol (21.2 mmol),3.58 g of 10-undecyn-1-ol (21.2 mmol) and 5.57 g of triphenylphosphine(21.2 mmol) with stirring in 40 mls of tetrahydrofuran (THF) at roomtemperature under argon. A solution of 3.7 g diethyl azodicarboxylate(21.2 mmol) was added dropwise to the reaction mixture at roomtemperature. The solution was allowed to stand overnight. The reactionmixture was concentrated, dissolved in ether, filtered and concentrated.This yielded 3.5 g of the acetylene product having the following formula##STR28## A 1 g portion of the above acetylene product (4.09 mmol) wasdissolved in 25 ml of THF and 10 mg of triphenyl methane was added as anindicator. The solution was cooled to -30° C. and 2.6 ml of 1.6M n-butyllithium (4.1 mmol) was added dropwise until the solution turned red. Thesolution was back-titrated with the acetylene product until colorless.The solution was cooled to -78° C. and 2 ml of trimethylsilyl chloride(15.8 mmol) was added. The solution was slowly warmed to roomtemperature. The reaction was quenched with water and extracted withhexane. The hexane extract was washed once with water and once withbrine and dried over MgSO₄. The above product was recovered in an amountof 1.26 g.

EXAMPLE 21 ##STR29##

The above compound is prepared by reacting 3 mmoles of the acid chlorideproduct from Example 1 with 0.95 g (3 mmoles) of the TMS-acetyleneproduct from Example 20. The acid chloride and the TMS-acetylene productare dissolved in 10 cc of dichloromethane and cooled to 0° C. To thereaction mixture is added 0.8 g (6 mmoles) of aluminum chloride (AlCl₃)in small portions over ten minutes. The reaction mixture is stirred forabout 1.5 hours at 0° C. The reaction is quenched with ice and themixture is extracted three times with diethyl ether. The extracts arecombined and washed once with water and once with brine (saturated NaCl)and dried over magnesium sulfate. Removal of the solvent in vacuo yieldsthe above product.

EXAMPLE 22 4-(1,3-dioxo-12-phenoxydodecyl)benzoic acid ##STR30##

The product of Example 21, 27 mg is treated with 2 ml of cold H₂ SO₄which has been cooled in an ice bath. The mixture is stirred until allof the product has dissolved. To the reaction mixture is added 1 mg HgO.The reaction mixture is maintained cool in an ice bath. Two drops ofwater are added to the reaction mixture. The ice bath is removed and thereaction mixture is stirred and allowed to warm to room temperature forone half hour. The mixture is cooled to 0° C. and quenched with ice. Thereaction mixture is diluted with water and extracted twice with ethylacetate. The collected extracts are washed twice with water. The washedethyl acetate fractions are collected and the volatile componentsremoved and the residue chromatographed on silica which yields the aboveproduct.

EXAMPLE 23 ##STR31##

The above acid chloride was prepared from isophthalic acid by reacting0.5 g (3 mmoles) of isophthalic acid with 2 cc of [COCl]₂ (23.6 mmoles)in 10 cc of benzene and with one drop of dimethylformamide. The reagentswere mixed and warmed to 60° C. for twenty-four hours. The reactionmixture was cooled to room temperature and the volatile components wereremoved in vacuo to give the above compound.

EXAMPLE 24

    CH.sub.3 O--(CH.sub.2).sub.9 --C.tbd.C--TMS

The above product was prepared by placing 1 g of hydroxy acetylene (5.94mmol) in 10 ml of THF and cooling to 0° C. To the solution was added 1ml of methyl iodide which was followed by the portionwise addition of0.4 g of sodium hydride (8.33 mmol). The reaction mixture was stirredand warmed to room temperature overnight. The reaction was quenched bypouring in to 100 ml of water, then extracted with hexane. The hexanewas washed once with water and once with brine and dried over magnesiumsulfate. Removal of the solvent in vacuo yielded 1.21 g of a pale yellowoil acetylene product of the following formula

    CH.sub.3 O--(CH.sub.2).sub.9 --C.tbd.CH

A 1 g portion of the above acetylene product (5.5 mmol) was dissolved in25 ml of THF and 10 mg of triphenyl methane was added as an indicator.The solution was cooled to -30° C. and 3.4 ml of 1.6M n-butyl lithium(4.1 mmol) was added dropwise until the solution turned red. Thesolution was back-titrated with the acetylene product until colorless.The solution was cooled to -78° C. and 2 ml of trimethylsilyl chloride(15.8 mmol) was added. The solution was slowly warmed to roomtemperature. The reaction was quenched with water and extracted withhexane. The hexane extract was washed once with water and once withbrine and dried over MgSO₄. The above product was recovered in an amountof 1.26 g.

EXAMPLE 25 ##STR32##

The above compound is prepared by reacting 3 mmoles of the acid chlorideproduct from Example 23 with 0.76 g (3 mmoles) of the TMS-acetyleneproduct from Example 24. The acid chloride and the TMS-acetylene productare dissolved in 10 cc of dichloromethane and cooled to 0° C. To thereaction mixture is added 0.8 g (6 mmoles) of aluminum chloride (AlCl₃)in small portions over ten minutes. The reaction mixture is stirred forabout 1.5 hours at 0° C. The reaction is quenched with ice and themixture is extracted three times with diethyl ether. The extracts arecombined and washed once with water and once with brine (saturatedsodium bicarbonate solution) and dried over magnesium sulfate. Removalof the solvent in vacuo yields 0.29 g of the above product.

EXAMPLE 26 3-(12-methoxy-1,3-dioxododecyl)benzoic acid ##STR33##

The product of Example 25, 27 mg is treated with 2 ml of cold H₂ SO₄which has been cooled in an ice bath. The mixture is stirred until allof the product has dissolved. To the reaction mixture is added 1 mg HgO.The reaction mixture is maintained cool in an ice bath. Two drops ofwater are added to the reaction mixture. The ice bath is removed and thereaction mixture is stirred and allowed to warm to room temperature forone half hour. The mixture is cooled to 0° C. and quenched with ice. Thereaction mixture is diluted with water and extracted twice with ethylacetate. The collected extracts are washed twice with water. The washedethyl acetate fractions are collected and the volatile componentsremoved and the residue chromatographed on silica gel which yields theabove product.

EXAMPLE 27 ##STR34##

The above product is prepared by reacting 1.9 g (10 mmol) ofp-hexoxyphenol, 2.1 g (20 mmol) 5-chloropentyne, 2.1 g potassiumcarbonate (15 mmol) in 50 ml of DMF, 100 mg of sodium iodide and heatingto 40° C. for 16 hours. The reaction mixture is poured into water andextracted with hexane. The organic extract is washed once with 10% NaOH,once with water and once with brine and dried over magnesium sulfate.Removal of the solvent yields a gum. The gum is purified by columnchromatography on silica to give the above acetylene product.

EXAMPLE 28 ##STR35##

A 1 g portion of the acetylene product (3.8 mmol) of Example 27 isdissolved in 25 ml of THF and 10 mg of triphenyl methane is added as anindicator. The solution is cooled to -30° C. and 3.4 ml of 1.6M n-butyllithium (4.1 mmol) is added dropwise until the solution turns red. Thesolution is back-titrated with the acetylene product until colorless.The solution is cooled to -78° C. and 2 ml of trimethylsilyl chloride(15.8 mmol) is added. The solution is slowly warmed to room temperature.The reaction is quenched with water and extracted with hexane. Thehexane extract is washed once with water and once with brine and driedover MgSO₄. After removal of the solvent the above product is recovered.

EXAMPLE 29 ##STR36##

The above compound is prepared by reacting 3 mmoles of the acid chlorideproduct from Example 1 with 1.0 g (3 mmoles) of the TMS-acetyleneproduct from Example 28. The acid chloride and the TMS-acetylene productare dissolved in 10 cc of dichloromethane and cooled to 0° C. To thereaction mixture is added 0.8 g (6 mmoles) of aluminum chloride (AlCl₃)in small portions over ten minutes. The reaction mixture is stirred forabout 1.5 hours at 0° C. The reaction is quenched with ice and themixture is extracted three times with diethyl ether. The extracts arecombined and washed once with water and once with brine (saturated NaCl)and dried over magnesium sulfate. Removal of the solvent in vacuo yieldsthe above product.

EXAMPLE 30 4-[6-[4-(hexyloxy)phenoxy]-1,3-dioxohexyl]benzoic acid##STR37##

The product of Example 29, 27 mg is treated with 2 ml of cold H₂ SO₄which has been cooled in an ice bath. The mixture is stirred until allof the product has dissolved. To the reaction mixture is added 1 mg HgO.The reaction mixture is maintained cool in an ice bath. Two drops ofwater are added to the reaction mixture. The ice bath is removed and thereaction mixture is stirred and allowed to warm to room temperature forone half hour. The mixture is cooled to 0° C. and quenched with ice. Thereaction mixture is diluted with water and extracted twice with ethylacetate. The collected extracts are washed twice with water. The washedethyl acetate fractions are collected and the volatile componentsremoved and the residue chromatographed on silica which yields the aboveproduct.

EXAMPLE 31 ##STR38##

The above acid chloride was prepared from phthalic acid by reacting 0.5g (3 mmoles) of phthalic acid with 2 cc of [COCl]₂ (23.6 mmoles) in 10cc of benzene and with one drop of dimethylformamide. The reagents weremixed and warmed to 60° C. for twenty-four hours. The reaction mixturewas cooled to room temperature and the volatile components were removedin vacuo to give the above compound.

EXAMPLE 32

    TMS--C.tbd.C--(CH.sub.2).sub.4 CF.sub.3

The above compound is prepared by first preparing a1-trifluoro-5-bromopentane by the reaction of 5-bromopentanoic acid withSF₄. The reaction is conducted by mixing 50.166 g (0.2771 mmol) of the5-bromopentanoic acid with 120 g (1.111 mmol) of SF₄ and heating in apressure vessel. A 12.5 g (0.3 mmol) amount of NaF was added in 50 mlmethylene chloride. The reaction mixture was filtered and washed withabout 60 ml of methylene chloride. The methylene chloride was washedonce with water and once with NaHCO₃ and once with brine and dried overmagnesium sulfate. Removal of the solvent in vacuo gave a brown oilwhich was distilled and the 25°-30° C. fraction collected yielding 42.73g of 1-trifluoro-5-bromopentane. Trimethylsilyl-acetylene, 5 g (50.9mmol) was mixed with 50 mg of triphenylmethane in 200 ml of THF andcooled to -50° C. and 1.6M n-butyl lithium in hexane was added dropwise(about 33 ml) until a red color persisted. A small portion oftrimethylsilyl acetylene was added until the red color disappeared. Themixture was warmed to -20° C. and stirred for one half hour. Thereaction mixture was cooled to -40° C. and 10 g of thetrifluoro-5-bromopentane was added dropwise. Then 50 ml of HMPA wasadded dropwise and the reaction mixture was stirred and warmed to roomtemperature. The reaction was quenched with water and was poured intoone liter of hexane. The mixture was washed four times with water, oncewith brine and dried over magnesium sulfate. The solvent was stripped invacuo to give a brown liquid that was distilled. The fraction boilingbetween 65°-68° C. was collected and gave 5.0 g of the above TMSacetylene product.

EXAMPLE 33 ##STR39##

The above compound is prepared by reacting 3 mmoles of the acid chlorideproduct from Example 31 with 0.66 g (3 mmoles) of the TMS-acetyleneproduct from Example 32. The acid chloride and the TMS-acetylene productare dissolved in 10 cc of dichloromethane and cooled to 0° C. To thereaction mixture is added 0.8 g (6 mmoles) of aluminum chloride (AlCl₃)in small portions over ten minutes. The reaction mixture is stirred forabout 1.5 hours at 0° C. The reaction is quenched with ice and themixture is extracted three times with diethyl ether. The extracts arecombined and washed once with water and once with brine (saturated NaCl)and dried over magnesium sulfate. Removal of the solvent in vacuo yieldsthe above product.

EXAMPLE 34 2-(8,8,8-trifluoro-1,3-dioxooctyl)benzoic acid ##STR40##

The product of Example 33, 27 mg is treated with 2 ml of cold H₂ SO₄which has been cooled in an ice bath. The mixture is stirred until allof the product has dissolved. To the reaction mixture is added 1 mg HgO.The reaction mixture is maintained cool in an ice bath. Two drops ofwater are added to the reaction mixture. The ice bath is removed and thereaction mixture is stirred and allowed to warm to room temperature forone half hour. The mixture is cooled to 0° C. and quenched with ice. Thereaction mixture is diluted with water and extracted twice with ethylacetate. The collected extracts are washed twice with water. The washedethyl acetate fractions are collected and the volatile componentsremoved and the residue chromatographed on silica which yields the aboveproduct.

EXAMPLE 35 ##STR41##

To a reaction vessel was added m-iodobenzoic acid (1 g, 4.03 mmoles)0.83 g acetylene (4.0 mmoles) of the formula H--C.tbd.C--C₁₃ H₂₇, 46 mgof a palladium catalyst Pd (PPh₃)₄ (0.04 mmoles) in 10 ml ofdiethylamine. The reaction vessel was degassed with argon. To thereaction vessel was added 0.15 g of copper iodide (0.8 mmoles). Thereaction mixture was stirred under argon for 21/2 days. The volatilecomponents were removed in vacuo and the residue was treated with 10%hydrochloric acid. The aqueous solution was extracted with ethyl acetateand the extract was washed with 10% hydrochloric acid, water and brine.The extract was dried over magnesium sulfate. The solvent was removed invacuo to give a brown gummy solid. This residue was chromatographed onsilica eluting with 10% ethyl acetate-90% hexane-1% acetic acid to yield1.19 g (3.6 mmoles) of a white solid of the above identified compound.

Analysis Calculated: C, 80.44; H, 9.82 Found: C, 80.65; H, 10.08

EXAMPLE 36 ##STR42##

The compound of Example 35, 0.33 g (1 mmole) was taken up in a solventmixture consisting of 4.6 ml of CCl₄, 4.6 ml of CH₃ CN and 7 ml of waterfollowed by addition of NaIO₄ (0.88 g, 4.1 mmoles) which was added inone portion. The reaction mixture was stirred for 10 minutes until aclear solution (two phase) formed. To the solution was added 0.0029 g(0.022 mmoles) of RuO₂. The reaction mixture was stirred for about 18hours at room temperature. The reaction mixture was transferred to aseparatory funnel and the organic layer separated. The aqueous layer wasextracted with dichloromethane. The combined extracts were dried overmagnesium sulfate and filtered through celite. Evaporation of thesolvent followed by chromatography on silica eluting with 2:1hexane-diethylether (1% HOAc) yielded 0.12 g of the above identifiedcompound.

Calculated: C, 73.30; H, 8.95 Found: C, 73.11; H, 8.94

EXAMPLE 37 ##STR43##

To a reaction vessel was added 3 g of m-iodobenzoic acid (12.1 mmoles)and 1.53 g of acetylene (15 mmoles) of the formula Ph--C.tbd.C--H. Tothe reaction vessel was added a palladium catalyst 0.085 g (0.121 mmolesof Pd(PPh₃)₂ Cl₂ in 30 ml of diethylamine. The reaction mixture washeated in an oil bath to 90° C. for 2 hours. The reaction mixture wascooled to room temperature. The volatile components were removed invacuo leaving a residue of a dark red solid. The residue wascrystallized from diethyl ether and hexane and triturated with hexane togive 1.64 g of a yellow solid of the above structure.

Analysis Calculated: Calculated for 0.1H₂ O: C, 80.41; H, 4.59 Found: C,80.49; H, 4.77

EXAMPLE 38 ##STR44##

The product of Example 37, 0.22 g (1 mmole) was taken up in a solventconsisting of 4.6 ml of CCl₄, 4.6 ml of CH₃ CN and 7 ml of water towhich was added NaIO₄ (0.88 g, 4.1 mmoles). The reaction mixture wasstirred for 10 minutes until a clear solution (two phase) developed. Tothe reaction mixture was added 0.0029 g (0.022 mmoles) of RuO₂. Thereaction mixture was stirred for approximately 18 hours at roomtemperature. The mixture was transferred to a separatory funnel and theorganic layer was separated. The aqueous layer was extracted withdichloromethane. The combined extracts were dried over MgSO₄ andfiltered through celite. Evaporation of the solvent followed bychromatography on silica eluting with 2:1 hexane/diethyl ether (1% HOAc)yielded 0.156 g of the above identified compound.

Analysis Calculated for 0.1H₂ O: C, 70.37; H, 4.02 Found: C,70.23;H,4.07

ASSAY FOR LTB₄ AND PGE₂ PRODUCTION BY HL-60 CELLS

HL-60 cells were induced to differentiate into granulocytes by a 4 dayincubation with 0.8% (v/v) N,N-dimethyl formamide as disclosed inFontana et al., Proc. Natl. Acad. Sci. 78 (6):3863-3866 (1981); Agins etal., Biochem. Biophys. Res. Comm. 126, 143-149 (1985); and Bonser etal., Biochemistry 20: 5297-5301 (1981). Prior to performing the assay,differentiated HL-60 cells were washed once with Hanks' balanced saltsolution containing 0.35 mg/ml sodium bicarbonate and 10 mM HEPES pH7.35 (HBSS). HL-60 cells (3×10⁶ cells/ml) were pre-incubated with thecompound tested or a control vehicle at 37° C. for 10 minutes, followedby 5 minute incubation with 5×10⁻⁶ M calcium ionophore A23187 in a finalvolume of 1.0 ml. After incubation, the cells were pelleted bycentrifugation and the LTB₄ and PGE₂ in the supernatent were quantifiedby radioimmunoassay. IC₅₀ values (means ±S.E.) for compounds herein thatwere tested are shown in the following Table and represent theconcentrations of the compound required to inhibit 50% of LTB₄ or PGE₂production by HL-60 cells stimulated with the calcium ionophore A23187.

HUMAN SYNOVIAL FLUID PHOSPHOLIPASE A₂ (HSF-PLA₂) ASSAY

Human synovial fluid phospholipase A₂ was purified approximately 5000fold following the procedures of Franson et al., Lung 160, 275-284(1982) and Fawzy et al., Bio Phys. J. 49, 533a (1986). Followingpurification the enzyme activity was measured by established methodologyusing [¹⁴ C]-oleate-labeled, autoclaved E. coli as the substrate as alsoshown in the above noted references. The assay was performed in a finalvolume of 100 μl containing 50 mM HEPES (pH 7.0), 150 mM NaCl, 5 mMCaCl₂, 7 nM [¹⁴ C]-oleate-labeled E. coli phospholipid and with orwithout the compound from one of the examples herein undergoing anassay. The compound or control vehicle was pre-incubated with the PLA₂for 5 minutes followed by addition of the E. coli substrate to initiatethe reaction. The reaction was maintained at 37° C. for 30 minutes andthen terminated by the addition of 2 ml tetrahydrofuran (THF). Thereaction product, [¹⁴ C]-oleic acid, was extracted using a 1 ml BondElut-NH₂ Solid phase extraction column. The IC₅₀ value for the compound(mean ±S.E.) is given in the following Table and represents theconcentration of the compound required to inhibit 50% of the PLA₂activity.

                  TABLE                                                           ______________________________________                                                         LTB4 Biosynthesis                                                             inhibition cells IC50 μM                                  Example #                                                                             HSF-PLA2 IC50 μM                                                                          Human PMNs  HL60 cells                                 ______________________________________                                        5       1.7            1.2         1.5                                        4       90                                                                    9       2.7            3.5         4.7                                        19      48             3.0                                                    15      1.9            2.7                                                    22      16             2.7                                                    ______________________________________                                    

We claim:
 1. A pharmaceutical composition comprising a compound of theformula ##STR45## or a pharmaceutically acceptable salt thereof whereinX is --CH═CH--;wherein R¹ is --CO₂ R² ; wherein R² is hydrogen, alkyl of1 to 6 carbons or a pharmaceutically acceptable cation; wherein R is analkyl of from 1 to 20 carbons, --(CH₂)_(p) CF₃ or --(CH₂)_(q) R³ whereinR³ is alkoxy, phenoxy or alkoxy substituted phenoxy wherein the alkoxygroup has from 1 to 8 carbons; wherein p and q are integers from 0 to20; wherein n is 1; and wherein m is 0, 1, 2, or 3; and apharmaceutically acceptable carrier.
 2. A method for the treatment ofmammals exhibiting an LTB₄ mediated inflammatory condition, comprisingadministering a compound of the formula ##STR46## or a pharmaceuticallyacceptable salt thereof wherein X is --CH═CH--;wherein R¹ is --CO₂ R² ;wherein R² is hydrogen, alkyl of 1 to 6 carbons or a pharmaceuticallyacceptable cation; wherein R is an alkyl of from 1 to 20 carbons,--(CH₂)_(p) CF₃ or --(CH₂)_(q) R³ wherein R³ is alkoxy, phenoxy oralkoxy substituted phenoxy wherein the alkoxy group has from 1 to 8carbons; wherein p and q are integers from 0 to 20; wherein n is 1; andwherein m is 0, 1, 2, or
 3. 3. A pharmaceutical composition according toclaim 1 wherein m is zero.
 4. A pharmaceutical composition according toclaim 1 wherein R¹ is --CO₂ H.
 5. A pharmaceutical composition accordingto claim 1 wherein R comprises an alkyl group of 10 to 15 carbons.
 6. Apharmaceutical composition according to claim 5 wherein the compound is3-(1,3-dioxohexadecanyl)benzoic acid.
 7. A pharmaceutical compositionaccording to claim 5 wherein the compound is4-(1,3-dioxohexadecanyl)benzoic acid.
 8. A method according to claim 2wherein m is zero.
 9. A method according to claim 2 wherein R¹ is --CO₂H.
 10. A method according to claim 2 wherein R comprises an alkyl groupof 10 to 15 carbons.
 11. A method according to claim 10 wherein thecompound is 3-(1,3-dioxohexadecanyl)benzoic acid.
 12. A method accordingto claim 10 wherein wherein the compound is4-(1,3-dioxohexadecanyl)benzoic acid.